A Stepwise, One-Pot, Multienzyme Cascade Reaction Coupled with Saccharomyces cerevisiae Used for the Biosynthesis of cis-Jasmone

cis-Jasmone is an essential compound used in perfumery, cosmetics, and food flavoring with distinct aromatic properties. Traditional production methods, such as plant extraction and chemical synthesis, have become increasingly challenged by resource and environmental concerns, while there is also a notable gap in the biotechnological approach toward the synthesis of cis-jasmone. Herein, we present a novel biosynthesis of cis-jasmone using a multienzyme cascade utilizing α-linolenic acid as a substrate to produce iso-12-oxophytodienoic acid via the sequential catalysis of lipoxygenase, Arabidopsis thaliana-derived allene oxide synthase and allene oxide cyclase 2, and Helicoverpa armigera-derived glutathione S-transferase 16, followed by its conversion to cis-jasmone by β-oxidation of Saccharomyces cerevisiae. After optimization of the cascade processes, cis-jasmone was successfully generated at 96 μg/L using a stepwise, one-pot process, which suggested the biosynthesis of cis-jasmone can be achieved in vitro. This biocatalytic approach fills the existing gap and offers a sustainable alternative to traditional jasmone production.